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Characterization and identification of fungicide insensitive Pestalotiopsis-like species pathogenic to tea crop in India

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Abstract

Gray blight, a fungal disease caused by Pestalotiopsis-like species, is a widespread disease affecting tea crop (Camellia sinensis (L.) Kuntze) in many tea-growing countries, including India, resulting in huge losses in tea production. In India, several studies have been conducted to understand the fungal diseases of tea crop, but gray blight has not been well described in major tea growing areas such as in North Bengal, based on its geographic distribution, molecular analysis, or pathogenicity, and even fungicide resistance. The objective of this study was to identify and characterize the causative agents of gray blight disease in symptomatic leaf sample of tea crop collected from 27 tea gardens located in North Bengal, India and to evaluate some common fungicides against them in order to understand the resistance mechanism. In this study, we characterized Pestalotiopsis-like species based on the phylogenies of DNA sequences (internal transcribed spacers) and assessment of conidial characteristics. The study revealed that out of 27 isolates of gray blight pathogens, 17 belonged to the genus Pseudopestalotiopsis (Ps.), six isolates were Neopestalotiopsis, and four were Pestalotiopsis. Two novel species, Ps. thailandica and N. natalensis were introduced through this study. The most frequently isolated genus from C. chinensis was Pseudopestalotiopsis. Pathogenicity tests showed that the isolates displayed significantly different virulence when inoculated onto wounded tea leaves and the mycelial growth rate was positively correlated with pathogenicity (P < 0.01). Based on the 13 ISSR (Inter Simple Sequence Repeat) markers used and principal coordinate analysis, it was found that isolates were very diverse. Out of 27 isolates, IND0P2, DLG0P10, and BHAT0P11 isolates were insensitive against both MBC + M3 (Carbendazim + Mancozeb) and DMI (Hexaconazole) fungicides, while isolates SANY0P18, PAHG0P19, RANG0P24, and SING0P25 were insensitive only against MBC + M3 fungicide. Further, these insensitive isolates were grouped into separate clusters by ISSR, indicating their distinctiveness. However, all the evaluated isolates were susceptible to M1 (copper oxychloride) and another DMI (propiconazole) fungicides. Therefore, to manage gray blight, fungicide resistance management strategies as recommended by Fungicide Resistance Action Committee should be implemented.

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References

  • Ali EM, Amiri A (2018) Selection pressure pathways and mechanisms of resistance to the demethylation inhibitor difenoconazole in Penicillium expansum. Front Microbiol 9:2472

    Google Scholar 

  • Baraldi E, Mari M, Chierici E, Pondrelli M, Bertollini P, Pratella GC (2003) Studies of thiabendazole resistance of Penicillium expansum of pears: pathogenic fitness and genetic characterization. Plant Pathol 52:362–370

    CAS  Google Scholar 

  • Bora P, Bora LC, Bhuyan RP, Hashem A, Abd-Allah EF (2022) Bioagent consortia assisted suppression in grey blight disease with enhanced leaf nutrients and biochemical properties of tea (Camellia sinensis). Biol Contr 170:104907

    CAS  Google Scholar 

  • Brent KJ, Hollomon DW (2007) Fungicide resistance: the assessment of risk, 2nd edn. Fungicide Resistance Action Committee, Brussels

    Google Scholar 

  • Chen Y, Zeng L, Shu N, Wang H, Tong H (2017) First report of Pestalotiopsis camelliae causing gray blight disease on Camellia sinensis in China. Plant Dis 101:1034

    Google Scholar 

  • Chen YJ, Zeng L, Shu N, Wang H, Tong HR (2018) Occurrence of Pestalotiopsis lushanensis causing gray blight disease on Camellia sinensis in China. Plant Dis 102:2654

    Google Scholar 

  • Chen Y, Wan Y, Zeng L, Meng Q, Yuan L, Tong H (2021) Characterization of Pestalotiopsis chamaeropis causing gray blight disease on tea leaves (Camellia sinensis) in Chongqing, China. Canadian J Plant Pathol 43:413–420

    CAS  Google Scholar 

  • Damicone JP, Smith DL (2009) Fungicide resistance management. Oklahoma State University Division of Agricultural Sciences and Natural Resources website. 23: Available: http://pods.dasnr.okstate.edu/docushare/dsweb/Get/Rendition-3508/F-7663web.pdf. Accessed 2022 March

  • Darapanit A, Boonyuen N, Leesutthiphonchai W, Nuankaew S, Piasai O (2021) Identification, pathogenicity and effects of plant extracts on Neopestalotiopsis and Pseudopestalotiopsis causing fruit diseases. Sci Rep 11:22606

    CAS  Google Scholar 

  • Fan J, Urban M, Parker JE, Brewer HC, Kelly SL, Hammond-Kosack KE, Fraaije BA, Liu X, Cools HJ (2013) Characterization of the sterol 14α-demethylases of Fusarium graminearum identifies a novel genus-specific cyp51 function. New Phytol 198:821–835

    CAS  Google Scholar 

  • Fan J, Luo Y, Michailides TJ, Guo L (2014) Simultaneous quantification of alleles E198A and H6Y in the β-tubulin gene conferring benzimidazole resistance in Monilinia fructicola using a duplex real-time (TaqMan) PCR. Pest Manag Sci 70:245–251

    CAS  Google Scholar 

  • Felsenstein J (1985) Confidence limits on phylogenies: An approach using the bootstrap. Evolution 39:783–791

    Google Scholar 

  • Fungicide Resistance Action Committee. 2022. Fungal control agents sorted by cross-resistance pattern and mode of action (including coding for FRAC Groups on product labels): Online publication: frac-code-list-2022--final.pdf

  • Gisi U, Chin KM, Knapova G, Küng Färber R, Mohr U, Parisi S, Sierotzki H, Steinfeld U (2000) Recent developments in elucidating mode of resistance to phenylamide, DMI, and strobilurin fungicides. Crop Prot 19:863–872

    CAS  Google Scholar 

  • Grover RK, Moore JD (1962) Toximetric studies of fungicides against brown rot organism- Sclerotinia fructicola and S. laxa. Phytopathol 52:876–880

    CAS  Google Scholar 

  • Gullino ML, Tinivella F, Garibaldi A, Kemmitt GM, Bacci L, Sheppard BL (2010) Mancozeb: past, present, and future. Plant Dis 94:1076–1086

    CAS  Google Scholar 

  • Haq UL, Ijaz I (2019) Assessment of genetic diversity based on ISSR markers in Neopestalotiopsis species collected from guava (Psidium guajava L.) plants affected with canker disease in Pakistan. Appl Ecol Env Res 17:11803–11811

    Google Scholar 

  • Hlaiem S, Yangui I, Rocca GD, Sara B, Danti R, Jamaa MLB (2022) First report of Pestalotiopsis biciliata causing dieback on Quercus coccifera and Pistacia lentiscus in Tunisia. Canadian J Plant Pathol. https://doi.org/10.1080/07060661.2022.2032831

    Article  Google Scholar 

  • Horikawa T (1986) Yield loss of new tea shoots due to grey blight caused by Pestalotia longiseta Spegazzini. Bull Shizuoka Tea Exp Stn 12:1–8

    Google Scholar 

  • Hu HL, Jeewon R, Zhou DQ, Zhou TX, Hyde KD (2007) Phylogenetic diversity of endophytic Pestalotiopsis species in Pinus armandii and Ribes spp.: evidence from rDNA and β-tubulin gene phylogenies. Fungal Divers 24:1–22

    CAS  Google Scholar 

  • Joshi SD, Sanjay R, Baby UI, Mandal AKA (2009) Molecular characterization of Pestalotiopsis spp. associated with tea (Camellia sinensis) in southern India using RAPD and ISSR markers. Indian J Biotechnol 8:377–383

    CAS  Google Scholar 

  • Keith LM, Velasquez ME, Zee FT (2006) Identification and characterization of Pestalotiopsis spp. causing scab disease of guava, Psidium guajava, in Hawaii. Plant Dis 90:16–23

    CAS  Google Scholar 

  • Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874

    CAS  Google Scholar 

  • Liu AR, Chen SC, Wu SY, Xu T, Guo LD, Jeewon R, Wei JG (2010) Cultural studies coupled with DNA based sequence analyses and its implication on pigmentation as a phylogenetic marker in Pestalotiopsis taxonomy. Mol Phylogenet Evol 57:528–535

    CAS  Google Scholar 

  • Liu F, Hou L, Raza M, Cai L (2017) Pestalotiopsis and allied genera from Camellia, with description of 11 new species from China. Sci Rep 7:866

    Google Scholar 

  • Liu F, Bonthond G, Groenewald JZ, Cai L, Crous PW (2019) Sporocadaceae, a family of coelomycetous fungi with appendage-bearing conidia. Stud Mycol 92:287–415

    CAS  Google Scholar 

  • Lonergan E, Pasche J, Skoglund L, Burrows M (2015) Sensitivity of Ascochyta species infecting pea, lentil, and chickpea to boscalid, fluxapyroxad, and prothioconazole. Plant Dis 99:1254–1260

    CAS  Google Scholar 

  • Maharachchikumbura SSN, Guo LD, Chukeatirote E, Bahkali AH, Hyde KD (2011) Pestalotiopsis—morphology, phylogeny, biochemistry and diversity. Fungal Divers 50:167–187

    Google Scholar 

  • Maharachchikumbura SSN, Guo LD, Cai L, Chukeatirote E, Wu WP, Sun X, Crous PW, Bhat DJ, McKenzie EHC, Bahkali AH, Hyde KD (2012) A multi-locus backbone tree for Pestalotiopsis, with a polyphasic characterization of 14 new species. Fungal Divers 56:95–129

    Google Scholar 

  • Maharachchikumbura SSN, Hyde KD, Groenewald JZ, Xu J, Crous PW (2014) Pestalotiopsis revisited. Stud Mycol 79:121–186

    CAS  Google Scholar 

  • Maharachchikumbura SSN, Guo LD, Liu ZY, Hyde KD (2016) Pseudopestalotiopsis ignota and Ps. camelliae spp. nov. associated with grey blight disease of tea in China. Mycol Prog 15:22–28

    Google Scholar 

  • Moller EM, Bahnweg G, Sandermann H, Geiger HH (1992) A simple and efficient protocol for isolation of high molecular weight DNA from filamentous fungi, fruit bodies, and infected plant tissues. Nucleic Acids Res 20:6115–6116

    CAS  Google Scholar 

  • Morales-Rodríguez C, Dalla M, Pia VM, Vannini AA (2019) Pestalotiopsis biciliata, a new leaf pathogen of Eucalyptus spp. recorded in Italy. Forest Pathol 49:e12492–e12499

    Google Scholar 

  • Nozawa S, Yamaguchi K, van Hop D et al (2017) Identification of two new species and a sexual morph from the genus Pseudopestalotiopsis. Mycoscience 58:328–337

    Google Scholar 

  • Omatsu N, Tomihama T, Nonaka Y (2012) Incidence of strobilurin and benzimidazole resistant strains of Pestalotiopsis longiseta, causal agent of gray blight, and practical control in tea field of Kagoshima Prefecture. Jpn J Phytopathol 78:3–9

    CAS  Google Scholar 

  • Owati AS, Agindotan B, Pasche JS, Burrows M (2017) The detection and characterization of QoI-resistant Didymella rabiei causing ascochyta blight of chickpea in Montana. Front Plant Sci 8:1165

    Google Scholar 

  • Pallavi RV, Nepolean P, Balamurugan A, Jayanthi R, Beulah T, Premkumar R (2012) In vitro studies of biocontrol agents and fungicides tolerance against grey blight disease in tea. Asian Pac J Trop Biomed 2:S435–S438

    Google Scholar 

  • Pandey AK, Burlakoti RR, Kenyon L, Nair RM (2018) Perspectives and challenges for sustainable management of fungal diseases of mungbean [Vigna radiata (L.) R. Wilczek var. radiata]: a Review. Front Environ Sci. 6:53

    Google Scholar 

  • Pandey AK, Sinniah G, Babu A, Tanti A (2021) How the global tea industry copes up with fungal diseases-challenges and opportunities. Plant Dis 105:1868–1879

    Google Scholar 

  • Patel P, Rajkumar BK, Parmar P, Shah R, Krishnamurthy R (2018) Assessment of genetic diversity in Colletotrichum falcatum Went accessions based on RAPD and ISSR markers. J Gen Eng Biotechnol 16:153–159

    Google Scholar 

  • Plant Protection Code (2021) Policy on usage of Plant Protection Formulations in Tea Plantations of India, Version 13, Tea Board of India. Online Publication PPC_Version_13_pdf3115.pdf (teaboard.gov.in).

  • Reddy MS, Murali TS, Suryanarayanan TS, Rajulu MG, Thirunavukkarasu N (2016) Pestalotiopsis species occur as generalist endophytes in trees of Western Ghats forests of southern India. Fungal Ecol 24:70–75

    Google Scholar 

  • Rohlf FJ (2002) NTSYs pc: Numerical taxonomy system, Ver: 2.01. Enter publishing, Ltd. Setauket, NY.

  • Sain SK, Pandey AK (2018) Application of talc-based formulation of some Trichoderma harzianum isolates to control soilborne diseases of brinjal and okra. Proc Natl Acad Sci, India, Sect B Biol Sci 88:905–914

    Google Scholar 

  • Samarakoon MC, Hyde KD, Promputtha I, Hongsanan S, Ariyawansa HA, Maharachchikumbura SSN, Daranagama DA, Stadler M, Mapook A (2016) Evolution of Xylariomycetidae (Ascomycota: Sordariomycetes). Mycosphere 7:1746–1761

    Google Scholar 

  • Shahriar SA, Nur-Shakirah AO, Mohd MH (2022) Neopestalotiopsis clavispora and Pseudopestalotiopsis camelliae-sinensis causing grey blight disease of tea (Camellia sinensis) in Malaysia. Eur J Plant Pathol 162:709–724

    CAS  Google Scholar 

  • Solarte FA, Muñoz CG, Maharachchikumbura S, Álvarez E (2018) Diversity of Neopestalotiopsis and Pestalotiopsis spp., causal agents of guava scab in Colombia. Plant Dis 102:49–59

    Google Scholar 

  • Takeda Y (2002) Genetic analysis of tea gray blight resistance in tea plants. JARQ 36:143–150

    Google Scholar 

  • Tamura K, Nei M (1993) Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol Biol Evo 10:512–526

    CAS  Google Scholar 

  • Tejesvi MV, Nalini MS, Mahesh B, Prakash HS, Kini KR, Shetty HS, Subbiah V (2007) New hopes from endophytic fungal secondary metabolites. Bol Soc Quim Mex 1:19–26

    Google Scholar 

  • Thind TS, Hollomon DW (2017) Thiocarbamate fungicides: reliable tools in resistance management and future outlook. Pest Manag Sci 74:1547–1551

    Google Scholar 

  • Tsai I, Maharachchikumbura SSN, Hyde KD, Ariyawansa HA (2018) Molecular phylogeny, morphology and pathogenicity of Pseudopestalotiopsis species on Ixora in Taiwan. Mycol Prog 17:941–952

    Google Scholar 

  • Tsai I, Chung C, Lin S, Hung T, Shen T, Hu S, Hozzein WN, Ariyawansa HA (2021) Cryptic diversity, molecular systematics, and pathogenicity of genus Pestalotiopsis and allied genera causing gray blight disease of tea in Taiwan, with a description of a new Pseudopestalotiopsis species. Plant Dis 105:425–443

    CAS  Google Scholar 

  • Tzean SS, Tzeng KC, Chang CA, Tsay TT, Yen SF (2019) List of plant diseases in Taiwan. Bureau of Animal and Plant Health inspection and Quarantine, Council of Agriculture, Executive Yuan, Taipei. Taiwan.

  • Wang S, Mi X, Wu L, Zhang L, Wei C (2019a) Characterization and pathogenicity of Pestalotiopsis-like species associated with gray blight disease on Camellia sinensis in Anhui Province, China. Plant Dis 103:2786–2797

    CAS  Google Scholar 

  • Wang Y, Xiong F, Lu Q, Hao X, Zheng M, Wang L, Li N, Ding C, Wang X, Yang Y (2019b) Diversity of Pestalotiopsis-like species causing gray blight disease of tea plants (Camellia sinensis) in China, including two novel Pestalotiopsis species, and analysis of their pathogenicity. Plant Dis 103:2548–2558

    CAS  Google Scholar 

  • Wei JG, Xu T, Guo LD, Liu AR, Zhang Y, Pan XH, Xu JG, Guo T, Liu LD, Zhang AR (2007) Endophytic Pestalotiopsis species associated with plants of Podocarpaceae, Theaceae and Taxaceae in southern China. Fungal Divers 24:55–74

    CAS  Google Scholar 

  • Wei JG, Phan CK, Wang L, Xu T, Luo JT, Sun X, Guo LD (2012) Pestalotiopsis yunnanensis sp. nov., an endophyte from Podocarpus macrophyllus (Podocarpaceae) based on morphology and ITS sequence data. Mycol Prog 12:563–568

    Google Scholar 

  • Wei J, Phan C, Wang L, Xu T, Luo J, Sun X, Guo L (2013) Pestalotiopsis yunnanensis sp. nov., an endophyte from Podocarpus macrophyllus (Podocarpaceae) based on morphology and ITS sequence data. Mycol Progress 12:563–568

    Google Scholar 

  • White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. Pages 315–322 in: PCR Protocols. A guide to methods and applications. Innis MA, Gelfand DH, Sninsky JJ, White TS, eds. Academic Press, San Diego, CA.

  • Wise KA, Bradley CA, Pasche JS, Gudmestad NC, Dugan FM, Chen W (2008) Baseline sensitivity of Ascochyta rabiei to azoxystrobin, pyraclostrobin, and boscalid. Plant Dis 92:295–300

    CAS  Google Scholar 

  • Wise KA, Bradley CA, Pasche JS, Gudmestad NC (2009) Resistance to QoI fungicides in Ascochyta rabiei from chickpea in the Northern Great Plains. Plant Dis 93:528–536

    CAS  Google Scholar 

  • Xu C, Liang L, Li Y, Yang T, Fan Y, Xuejin F, Wang MY (2021) Studies of quality development and major chemical composition of green tea processed from tea with different shoot maturity. LWT 142:111055

    CAS  Google Scholar 

  • Yamada K, Sonoda R (2012) Characterization of moderate resistance to QoI fungicides in Pestalotiopsis longiseta and polymorphism in exon–intron structure of cytochrome b gene. J Gen Plant Pathol 78:398–403

    CAS  Google Scholar 

  • Yamada K, Sonoda R, Ishikawa K (2016) Population genetic structure of QoI-resistant Pestalotiopsis longiseta isolates causing tea gray blight. Plant Dis 100:1686–1692

    Google Scholar 

  • Yang H, Xue X, Li H, Apandi SN, Tay-Chan SC, Ong SP et al (2018) The relative antioxidant activity and steric structure of green tea catechins—a kinetic approach. Food Chem 257:399–405

    CAS  Google Scholar 

  • Yong YC, Chen YJ, Fang BY, Cheng WH (2014) Sensitivity of Pestalotiopsis spp. from guava to benzimidazoles in Taiwan. Plant Pathology Bulletin, cabdirect.org.

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Acknowledgements

Dr. Abhay K. Pandey is thankful for the Start-up Research Grant (SRG/2021/000299) obtained from Department of Science and Technology (DST), Science and Engineering Research Board, Government of India.

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Science and Engineering Research Board,SRG/2021/00299

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AKP: Conceptualization of idea, conducted experiments, original draft preparation and data analysis, MH: Molecular analysis of isolates, review and edit, Vandana: Molecular analysis experiments, review and edit, AB and PD: Review and edit.

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Correspondence to Abhay K. Pandey or Manjunath Hubbali.

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Pandey, A.K., Hubbali, M., Vandana et al. Characterization and identification of fungicide insensitive Pestalotiopsis-like species pathogenic to tea crop in India. World J Microbiol Biotechnol 39, 34 (2023). https://doi.org/10.1007/s11274-022-03474-3

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